Capillary cassette and method of manufacturing the same

ABSTRACT

Capillary columns ( 102 ) pass through and are inserted in a rubber plate ( 14 ), held and fixed by elastic force of rubber, and two-dimensionally arranged on a sample injection side. It fixes the capillary columns ( 102 ) arranged on a plane in close contact by holding the same with a holder plate ( 6   a ) from below and with a rubber plate ( 16 ) from above on a detection side. In order to press the capillary columns ( 102 ) against the holder plate  6   a  and fix the same with the rubber plate ( 16 ), a holder plate ( 6   b ) fixing the rubber plate ( 16 ) to the holder plate ( 6   a ) on both sides of the arrangement of the capillary columns ( 102 ) is provided.

[0001] This application is a division of U.S. application Ser. No.09/402,890, filed Oct. 14, 1999; which is a 35 U.S.C. 371 application ofinternational application no. PCT/JP99/00653 filed Feb. 15, 1999, whichis based on Japanese application nos. 10-51489, 10-51490, 10-51491, and10-51492, all filed Feb. 16, 1998.

TECHNICAL FIELD

[0002] The present invention relates to, in a multi-capillaryelectrophoretic apparatus comprising such a multi-capillary arraymigration part that a plurality of capillary columns are arranged sothat samples are injected into the respective capillary columns andsimultaneously electrophoresed in all capillary columns and an opticalmeasuring part irradiating capillaries with light in the multi-capillaryarray migration part and measuring absorbance by the samples in theirradiated parts and fluorescence from the samples, a capillary cassetteforming the multi-capillary array migration part.

[0003] Such a multi-capillary electrophoretic apparatus is used forseparating/analyzing nucleic acid, protein, peptide, sugar and the like,and particularly plays an important role for analysis of the basesequence of DNA. The multi-capillary electrophoretic apparatus forsequence determination of DNA employs Sanger's reaction, electrophoresesa DNA fragment (fragment) sample labeling a primer or a terminator witha fluorescent material and detects fluorescence from the DNA fragmentsample during migration for deciding the base sequence.

BACKGROUND TECHNIQUE

[0004] For sequence determination of DNA having long base sequence suchas a human genome, a DNA sequencer having high sensitivity, a high speedand large throughput is necessary. As one method thereof, amulti-capillary DNA sequencer arranging capillary columns charged withgels in plural is proposed in place of that employing flat plate typeslab gels. In the capillary column, a sample is not only easy to handleor inject but also can be migrated at a high speed and detected in highsensitivity as compared with the slab gel. Namely, such problems resultfrom influence by Joulean heat that a band spreads and a temperaturegradient takes place applying a high voltage in the slab gel, while suchproblems are less in the capillary column and spreading of a band issmall to allow high sensitivity detection even if making high-speedmigration while applying a high voltage.

[0005] When making electrophoresis employing a plurality of capillarycolumns, it is desirable that attachment/detachment of the capillarycolumns to/from an electrophoretic apparatus body is easy, and it isdesirable to cassette the same with a holder for that. In considerationof the recent environmental problem, it is also preferable that thecapillary columns can be readily separated from the holder whendiscarded after use.

[0006] Furthermore, since a step by pressurization or decompression isrequisite for charging gels into the capillary columns, airtightness isnecessary for fixation of the capillary columns to the holder at leaston sample injection sides in order to simultaneously charge the gelsinto all capillary columns contained in the cassette.

[0007] Accordingly, a first objective of the present invention is toprovide a capillary cassette which can readily separate capillarycolumns from a holder and has sufficient airtightness between thecapillary columns and the holder on sample injection sides.

[0008] The capillary columns are so thin that the same are difficult tomanually work and are easily breakable. When fixing the capillarycolumns to the holder, it is a difficult operation requiring skill tobundle the same in plural, and in addition, it requires a long time.

[0009] Accordingly, a second objective of the present invention is tomake it possible to simply and quickly perform preparation of acapillary cassette.

[0010] The capillary columns are protected with various coats, in orderto increase mechanical strength. For coats, polyimide, silicone resin,polytetrafluoroethylene, acrylic resin and the like are used. When usingthe same as coats for capillary columns of electrophoresis however,optical characteristics of the coats such as light absorption andfluorescence hinder detection since optical means for absorbancemeasurement and fluorescence measurement are employed for detection ofsamples migrating in the capillary columns. Therefore, when the coatsare made of materials hindering optical detection, the coats ofdetection parts are burned by flames and removed with burning means suchas a lighter or a gas burner.

[0011] A multi-capillary electrophoretic apparatus using a plurality ofcapillary columns and simultaneously detecting a plurality of samplesaligns, arranges and fixes such capillary columns that coats ofdetection parts are removed, thereby forming detection windows when thecoats are made of materials hindering optical detection.

[0012] While a number of capillary columns are arranged in themulti-capillary electrophoretic apparatus, it follows that whenperforming coat removal of the detection parts on every one of thecapillary columns, a long time is required for the operation.

[0013] Furthermore, the coating materials include such a one that a partlocated in a flame is completely burned and removed in the process ofremoving the coating material by burning by the flame while the same ismelted but does not come to be removed in a portion separate from theflame and convex parts resulting from solidification of the melted coatare formed on both sides of the part from which the coat is removed. Ifsuch convex portions are formed, adjacent capillary columns do notadhere to each other or come to different levels due to the convexportions and detection windows are not planarly aligned when arrangingthe capillary columns, aligning positions of the parts from which thecoats are removed and forming the detection windows. Consequently, thefocus of an optical system may so deviate in detection that it cannotperform proper detection.

[0014] Accordingly, a third objective of the present invention is, whena coat for a capillary column is made of a material hindering opticaldetection, to provide such a capillary cassette that a detection windowof a capillary column employed for a multi-capillary electrophoreticapparatus is planarly formed to be suitable for optical measurement anda method of manufacturing a capillary cassette having a feature in adetection window forming step thereof.

[0015] An initial capillary electrophoretic apparatus is that employinga single capillary column. In this case, it dips an end of the capillarycolumn into a gel solution in a vessel, closes the vessel andpressurizes the vessel thereby pushing the gel into the capillary columnand charging the same as a method of charging the gel into the capillarycolumn. As another method, an operation of dipping an end of thecapillary column into the gel solution, decompressing another end of thecapillary column and sucking the gel into the capillary column, therebycharging the same, is also performed.

[0016] In multi-capillary electrophoresis, it is necessary to aligncapillary column end surfaces on sample injection sides in sampleinjection while it is necessary to align and handle capillary coatremoving parts on detection sides when performing detection for handlinga plurality of capillary columns, and hence it is desirable to providesuch a capillary cassette that arrangement is fixed by a holder so thatboth the sample injection sides and the detection sides are inprescribed arrangement relation.

[0017] Furthermore, it is extremely troublesome to charge a gel intoevery one of the capillary columns in the state of the capillarycassette, and, in actuality, impossible in practice. Therefore, awaitedis means for making it possible to simply perform gel charging into allcapillary columns included therein in units of the capillary cassette.

[0018] Accordingly, a fourth objective of the present invention is toprovide an apparatus to make it possible to simply charge gels into allcapillary columns included in such a capillary cassette.

DISCLOSURE OF THE INVENTION

[0019] In order to attain the first objective, the present invention issuch that, in a capillary cassette two-dimensionally arranging andfixing a plurality of capillary columns used in a multi-capillaryelectrophoretic apparatus on sample injection sides by a holder whilearranging the same in a line on a plane on detection sides (sidesopposite to the sample injection sides), the holder comprises a rubberplate so that end portions of the plurality of capillary columns passthrough the rubber plate one by one to be fixed by elastic force of therubber plate.

[0020] By inserting the capillary columns into the rubber plate andfixing the same with the holder on the sample injection sides,airtightness between the rubber plate and the capillary columns can bemaintained and it is possible to serve a sealing function with respectto pressurization or suction in gel charging into capillaries. Thecapillary columns are not fixed by an adhesive or the like, whereby thecapillary columns can be readily extracted from the holder in disposalof the capillary columns or the like.

[0021] In the present invention, it is preferable to comprise anotherholder pressing and holding the capillary columns from at least singlesurface sides of the plurality of capillary columns arranged in a lineon the plane through the rubber plate on the detection sides of thecapillary columns. By thus making fixation on the detection sides bypushing the capillary columns through the rubber plate, it is easy todetach the capillary columns from the holder on the detection sides, andan operation of separating the capillary columns and the holder indisposal after use or exchanging defective capillary columns becomeseasy.

[0022] Furthermore, the holder on the detection sides preferablycomprises clamping means fixing the capillary columns on both sideportions while pressing and fixing the same on at least two portions ofan intermediate part as means fixing the plurality of capillary columnsarranged in a line on the plane through the rubber plate. By making itpossible to press the capillary columns also in at least two portions ofthe intermediate part, it becomes easy to maintain arrangement on theplane.

[0023] A capillary cassette preparation method according to the presentinvention to attain the second objective is a method of passing througha supporter of a prescribed fixing position of capillary column fixingmeans having the supporter consisting of a rubber plate with a needlehaving an inner diameter larger than a capillary column from itsvertical direction, guiding the capillary column into the needle andmaking the same pass through the supporter, thereafter extracting onlythe needle thereby holding and fixing the capillary column by thesupporter and repeating this operation while changing the fixingposition on the supporter thereby successively two-dimensionallyarranging and fixing a plurality of capillary columns.

[0024] The capillary column guided in the needle to pass through thesupporter of the rubber plate on a sample injection side holder(capillary column fixing means) is held and fixed by elastic force ofrubber after only the needle is extracted from the supporter.

[0025] A capillary cassette preparation apparatus according to thepresent invention for executing this capillary cassette preparationmethod comprises arrangement position decision means holding capillarycolumn fixing means having a supporter consisting of a rubber plate formoving and fixing the same in an in-plane direction of the supporter, aneedle passing through the supporter comprised in the capillary columnfixing means from a vertical direction of its plane and guiding acapillary column to the fixing position, a first capillary column holderholding the capillary column and inserting the same into the needle, aguide guiding the forward end of the capillary column held by the firstcapillary column into the needle, slide guide means moving the needle,the guide and the first capillary column holder in a rectilineardirection perpendicular to the plane of the supporter, a secondcapillary column holder holding the forward end portion of the capillarycolumn made to project from the forward end of the needle in such astate that the needle passes through the supporter of the capillarycolumn fixing means, cut means cutting the capillary column in aprescribed length, and a roller unit successively feeding the capillarycolumn in an insert direction.

[0026] The fixing position of the capillary column is decided bymounting the capillary column fixing means on the arrangement positiondecision means and moving the arrangement position decision meansupward, downward, leftward and rightward. It linearly moves the needlein the capillary column fixing means direction along the slide guidemeans and makes the same pass through the supporter on the fixingposition. The forward end of the capillary column wound on a capillarydrum is guided into the needle by the guide following the needle alongthe slide guide means and the first capillary column holder, andprojects from the forward end of the needle. It holds the forward endportion of the capillary column by the second capillary column holdercomprised on a needle projection side of the arrangement positiondecision means, and extracts only the needle from the supporter of thecapillary column fixing means. The capillary column forward end portionremains in the supporter, elastic force of rubber which is the supporteracts in a direction closing a hole opened by the needle, and thecapillary column is fixed to the capillary column fixing means. It cutsthe fixed capillary column in the prescribed length by the cut means.The arrangement position decision means moves and brings a next fixingposition onto a moving straight line of the needle, the guide and thecapillary column holder. The capillary column wound on the drum istimely fed by the roller unit having a motor for motive power.

[0027] According to this capillary cassette preparation apparatus,capillary columns of a predetermined length are aligned in determinedorder by operating a lever in accordance with a prescribed procedure sothat a capillary cassette can be prepared, whereby the efficiency of apreparation operation for the capillary cassette remarkably rises.Furthermore, it can simply make transition to automatization by mountinga driving source, whereby simple preparation of a capillary cassettewith less failure is enabled.

[0028] In order to also form detection windows simultaneously withcapillary cassette preparation, it is preferable to further comprisedetection window preparation means removing capillary column coats ofprescribed positions from the forward ends of the capillary columns andpreparing the detection windows and arrangement means arrangingterminating end sides of the cut capillary columns in a line indetermined order in the capillary cassette preparation apparatus.

[0029] After the capillary columns are guided in the needle and passthrough the capillary column fixing means, a voltage is applied to thedetection window preparation means to prepare the detection windows onprescribed positions from the forward ends of the capillary columns.Thereafter the terminating ends of the capillary columns cut in theprescribed length are successively dropped between planes and arrangedin a line by the arrangement means formed by two planes having aparallel clearance slightly wider than the capillary column outerdiameter.

[0030] A capillary cassette according to the present invention forattaining the third object is such a one that a plurality of capillarycolumns are fixed by a holder on sample injection sides and planarlyarranged in a line on detection sides, coats of the capillary columnsare removed so that strip detection windows extending in the capillarycolumn arrangement direction are formed on the capillary columnarrangement on the detection sides, the capillary columns adhere to eachother and the coats fuse to each other so that the capillary columnarrangement is integrated around the detection windows.

[0031] In this capillary cassette, the capillary columns adhere to eachother, the coats fuse to each other and the capillary column arrangementis integrated around the detection windows, whereby the detectionwindows planarly align, the focus of an optical system in detection doesnot deviate, and it is possible to perform correct detection.

[0032] When manufacturing the capillary cassette, a manufacturing methodaccording to the present invention forms the detection windows includingthe following steps (A) and (B):

[0033] (A) a step of fixing the sample injection sides of the capillarycolumns using the holder and thereafter arranging and holding thecapillary columns in a line planarly in close contact with each other,and

[0034] (B) a step of bringing heating means having a length for aplurality of capillary column outer diameters into contact with orapproximating the same to a detection window formation planned region ofthe held capillary column arrangement for removing coats of theplurality of capillary columns while melting the coats around regionsfrom which the coats are removed and thereafter solidifying the same formaking the coats of the adjacent capillary columns fuse to each other.

[0035] It planarly arranges and fixes the capillary columns notsubjected to coat removal of detection parts in a line andsimultaneously performs coat removal of detection window formationplanned positions of the plurality of capillary columns with the heatingmeans. By simultaneously heating and removing the coats of the pluralityof capillary columns, the adjacent capillary columns are fused andintegrated with each other on positions separate from the regions fromwhich the coats are removed when the melted coats are cooled andsolidified. Thus, the plurality of capillary columns arranged in a lineform a flat cable.

[0036] For a means of heating, a nichrome wire heater or a ceramicheater can be employed.

[0037] When using as the heating means employed for removing the coatsthat whose length is shorter than the width of the capillary columnarrangement on the detection sides, it repeats the step (B) a number oftimes in the width direction of the capillary column arrangement forforming the detection windows.

[0038] This manufacturing method planarly arranges the plurality ofcolumns and thereafter simultaneously removes the coats of the detectionparts of the plurality of capillary columns, whereby remarkablereduction of the detection window preparation time can be made ascompared with the case of removing the coat as to every single capillarycollar.

[0039] In the peripheral parts of the detection windows formed by thismanufacturing method, the coats melted but not come to be removed fusethe adjacent capillary columns when cooled and solidified. The pluralityof fused capillary columns are previously planarly arranged and hencecome to planarly maintain the detection windows after fusion, and it ispossible to readily manufacture a capillary cassette capable ofperforming correct optical detection.

[0040] When fixing the detection sides of the plurality of capillarycolumns, it saves trouble of fixing the same one by one while it doesnot fix the same with an adhesive, whereby a time for drying theadhesive can also be saved.

[0041] A gel charging apparatus according to the present invention forattaining the fourth object is an apparatus for charging gels intocapillary columns of a capillary cassette. In the capillary cassette,end portions of sample injection sides of a plurality of capillarycolumns mounted on a multi-capillary electrophoretic apparatus aretwo-dimensionally arranged while passing through a holding member of aholder and fixed while keeping airtightness between the same and theholding member.

[0042] Gel charging into the capillary columns can be performed eitherby suction or pressurization. In gel charging, it does not seal therespective capillary columns one by one to perform suction orpressurization but seals all capillary columns of the capillary cassetteby the holder fixing the capillary columns while keeping airtightness tosimultaneously perform gel charging.

[0043] In the system performing gel charging by suction, the presentinvention comprises a chamber provided with an opening in its uppersurface so that closure means introducing end portions on sampleinjection sides of the capillary columns inside and closing the openingwith the holder is provided in the opening and further provided with anexhaust port and a release port to the atmosphere, a gel vessel storinga gel solution so that end portions of the capillary columns opposite tothe sample injection sides are dipped therein, exhaust means provided onthe exhaust port of the chamber, and a switching valve provided on therelease port.

[0044] When performing gel charging by suction, it dips the forward endsof the detection sides of all capillary columns of the capillarycassette in gels contained in the gel vessel, closes the opening of thechamber with the holder for the capillary cassette, decompresses thechamber with the exhaust means and inhales the gel solution into thecapillary columns.

[0045] In the system performing gel charging by pressurization, thepresent invention comprises a chamber provided with an opening on itsupper surface so that closure means introducing end portions of thesample injection sides of the capillary columns inside and closing theopening with the holder is provided in the opening while storing a gelsolution on such a position that the end portions on the sampleinjection sides of the capillary columns are dipped therein in such astate that the opening is closed with the holder and further providedwith a pressurization port and a release port to the atmosphere,pressurization means provided on the pressurization port of the chamberand a switching valve provided on the release port.

[0046] When performing gel charging by pressurization, it introduces thegel solution into the chamber, dips the forward ends of the capillarycolumns on the sample injection sides in the gel solution, closes theopening of the chamber with the holder, pressurizes the chamber with thepressurization means and pressure-fits gels into the capillary columns.

[0047] When charging the gel solution into the plurality of capillarycolumns forming a capillary array mounted on a multi-capillaryelectrophoretic apparatus, it does not directly close/fix the respectivecapillary columns but seals the plurality of capillary columns to theclosure means with the sample injection side holder fixing the same withexcellent airtightness to perform charging of the gels, whereby gelcharging for the plurality of capillary columns can be simultaneouslyperformed. Furthermore, simple mounting and airtightness in mounting canbe compatibly attained.

[0048] Charging of the gel solution into the plurality of capillarycolumns can be simultaneously performed, whereby it does not damage themaximum merit of improvement of throughput by simultaneous performanceof migration of a plurality of samples in multi-capillaryelectrophoresis in the process of gel preparation which is thepretreatment thereof.

[0049] It is preferable that a flow control valve controlling the flowrate is provided between the release port of the chamber and theswitching valve. While the pressure in the chamber is released to theatmosphere after gel charging into the capillary columns, the speed atwhich the internal pressure of the chamber returns can be controlledwith the flow control valve. While air may be mixed into the gelscharged into the capillary columns when the degree of decompression orpressurization in the chamber is large, the degree of decompression orpressurization can be controlled with the flow control valve in thiscase.

[0050] In the system performing gel charging by pressurization, the gelsolution may be directly introduced into the chamber, while it ispreferable in the aspect of maintenance of the chamber when providing anattachable/detachable vessel in the chamber for storing the gel solutionin the vessel.

BRIEF DESCRIPTION OF THE DRAWINGS

[0051]FIG. 1 is a schematic perspective view showing an example of amulti-capillary electrophoretic apparatus to which a capillary cassetteaccording to the present invention is applied.

[0052]FIG. 2 is a front elevational view of a capillary cassette of oneembodiment.

[0053]FIG. 3 is a left side elevational view of the capillary cassetteof the embodiment.

[0054]FIG. 4 is a top plan view of the capillary cassette of theembodiment.

[0055]FIG. 5 shows diagrams mounting a carrier on the embodiment, (A) isa front elevational view, (B) is a left side elevational view, and (C)is a top plan view.

[0056]FIG. 6 is a schematic perspective view of one embodiment of acapillary cassette preparation apparatus according to the presentinvention.

[0057]FIG. 7 is a plan view of the embodiment.

[0058]FIG. 8 is a front elevational view of the embodiment.

[0059]FIG. 9 is a flow chart of the operation procedure of theembodiment.

[0060]FIG. 10 is a schematic perspective view showing the final step ofone embodiment of a capillary cassette manufacturing method according tothe present invention.

[0061]FIG. 11 is a front elevational view showing a schematic structureof one embodiment of a capillary column gel charging apparatus accordingto the present invention in a partially fragmented manner.

[0062]FIG. 12 is a plan view showing the embodiment in a partiallyfragmented manner.

[0063]FIG. 13 is a sectional view showing the embodiment in a partiallyfragmented manner, and that showing a state cut on an A-A line positionof FIG. 12.

BEST MODES FOR CARRYING OUT THE INVENTION

[0064]FIG. 1 shows a schematic perspective view of an example of amulti-capillary electrophoretic apparatus to which a multi-capillarycassette according to the present invention is applied.

[0065] A capillary cassette 2 is such a one that a plurality ofcapillary columns 102 are arranged and fixed by holders 4 and 6 to forma capillary array, and an end 2 a of the capillary array defines asample injection side and is two-dimensionally arranged and fixed by theholder 4, to come into contact with a buffer solution of a reservoir 62for migration after sample injection. A terminating end 2 b of thecapillary array is such that the capillary columns 102 are arranged in aline on a plane, to come into contact with a buffer solution of areservoir 56 on the forward end. Such a detected portion 2 c that thecapillary columns 102 are arranged in a line and supported by the holder6 is provided on a terminating end side (detection side) of thecapillary array. The capillary columns 102 are coated with coats to beprotected against breakage. When a fluorescent detection method isemployed for detecting migrating samples and if the coats emitfluorescence, it removes the coats on the detected portion 2 c. Whenusing those coated with coats which are transparent and made of anon-fluorescent material, the coats of 102 of the capillary columns donot have be removed also on the detected portion 2 c. A number ofcapillary columns, e.g., 384 capillary columns are arranged on thecapillary cassette 2.

[0066] Different samples are injected into the respective capillarycolumns 102, and electrophoresis is simultaneously performed.

[0067] For an excitation light source 8 to be used for exciting thesamples themselves or fluorescent materials labeling the samples, anargon gas laser unit, for example, is provided. 10 is anexcitation-photoreceiving optical system, which is that irradiating thecapillary columns 102 of the detected portion 2 c with excitation lightand detecting fluorescence from the samples, and scanned by a scanningmechanism (illustration omitted) in a scan direction parallel to thearrangement plane of the capillary cassette in the detected portion 2 cand perpendicular to a migration direction. In order to make theexcitation light beam from the excitation light source 8 not deviateeven by scanning of the excitation-photoreceiving optical system 10, thelaser beam from the excitation light source 8 is guided to theexcitation-photoreceiving optical system 10 through optical fiber 9coupled by a coupler here as an example.

[0068] A migration buffer solution is contained in the lower reservoir62, the lower end 2 a of the capillary array is dipped in the buffersolution, and a migration voltage is applied to the capillary ends ofthe lower end 2 a of the capillary cassette 2 through the buffersolution. An upper electrode 54 is dipped in and comes into contact withthe buffer solution of the upper reservoir 56, a lower electrode 58 isdipped in and comes into contact with the buffer solution of the lowerreservoir 62, and the migration voltage is applied to both electrodes 54and 58 from a high pressure power source 60. Its power supply voltage is30 kV, for example, and a current capacity is 10 to 30 mA.

[0069] The migrated samples are protein samples, DNA fragments labeledwith a fluorescent material or the like.

[0070]FIG. 2, FIG. 3 and FIG. 4 show schematic block diagrams of acapillary cassette of one embodiment. FIG. 2 is a front elevationalview, FIG. 3 is a left side elevational view, and FIG. 4 is a top planview. The same reference numerals are assigned to parts playing the sameroles as FIG. 1.

[0071] A holder 4 on a sample injection side has a coat, and is such aone that a rubber plate 14 made of silicone rubber of 5 mm in thicknessholding and fixing capillary columns 102 with a hole is held betweenholder plates 4 a and 4 b of resin for two-dimensionally arranging thequartz glass capillary columns 102 of 300 μm in outer diameter and 100μm in inner diameter, for example, and integrated by fixing screws 4 c.384 holes passing the capillary columns 102 therethrough are provided onboth holder plates 4 a and 4 b in two dimensions of 16 by 24 of 4.5 mmpitches in correspondence to positions of respective holes of a 384-holemicroplate used for sample introduction. The holes of the holder plates4 a and 4 b are set to be larger than the outer diameters of thecapillary columns 102. The capillary columns 102 pass through bothholder plates 4 a and 4 b and the rubber plate 14 held therebetween andare held in the holes of the rubber plate 14 by elasticity of rubber,thereby being fixed while keeping airtightness between the same and theholder 4. A carrier fixture 20 is provided on an end of the holder plate4 a.

[0072] A holder 6 on a detection side fixes the capillary columns 102closely arranged on a plane by holding the same with a holder plate 6 afrom below and with a rubber plate 16 of silicone of 2 mm in thicknessfrom above. In order to press the capillary columns 102 against theholder 6 with the rubber plate 16 and fix the same, holder plates 6 bfixing the rubber plate 16 to the holder plate 6 a by fixing screws 6 care provided on both side portions of the arrangement of the capillarycolumns 102.

[0073] When arranging the 384 capillary columns 102 in a line in theholder 6, the width becomes about 12 cm even if arranging the same inclose contact, and hence the rubber plate 16 is bent outward at thecentral portion when fixing both end portions, and hence the force ofthe rubber plate 16 holding the capillary columns 102 weakens around thecenter of the holder 6. So that the rubber plate 16 does not float in adirection separating from the holder plate 6 a between both end portionsat which the rubber plate 16 is fixed to the holder plate 6 a by theholder plate 6 b, two grooves 17 a extending in the arrangementdirection of the capillary columns 102 are provided inside (the sideopposed to the rubber plate 16) of the holder plate 6 b, clamp bars 18 bpushing the rubber plate 16 in the direction of the holder plate 6 a areengaged in several portions, e.g., in positions of six portions in totalincluding three portions on either side, and a clamp screw 18 a fittedwith the holder plate 6 b to project outward from the holder plate 6 bis provided in each clamp bar 18 b. By adjusting the clamp screw 18 a,clamping force pressing the capillary columns 102 through the rubberplate 16 can be adjusted.

[0074] The total length of the capillary columns 102 is a 500 mm, andthe detected portion 2 c is provided on a position of about 400 mm froma sample injection end. In order to form detection windows on thedetected portion 2 c, a slot 17 c along the arrangement direction of thecapillary columns 102 is opened in the holder plates 6 a and 6 b and therubber plate 16 to define the detected portion 2 c. Signal detection inelectrophoresis is performed through the slot 17 c.

[0075] In order to fix the capillary columns 102 to the holder 4, itholds the rubber plate 16 between the holder plates 4 a and 4 b, passesthe rubber plate 14 with a needle along the opening holes of the holderplate 4 b, thereafter inserts the capillary columns 102 into the needle,and extracts only the needle, as described later with reference to FIG.6 to FIG. 9 in detail. Thus, the capillary columns 102 pass through therubber plate 14, and are supported and fixed by the rubber plate 14 in astate held by elastic force of rubber. Thereafter it cuts the capillarycolumns 102 in a prescribed length. By repeating this operation whilechanging the positions of the holes of the holder plate 4 b, thecapillary columns 2 are two-dimensionally arranged and fixed to theholder 4.

[0076] In this embodiment, the capillary columns 102 are fixed not onlyto the holder 4 but also to the holder 6 employing no adhesives.Therefore, the capillary columns 102 are readily separated from theholders 4 and 6, and it is also possible to exchange defective capillarycolumns.

[0077] Also, as described later, a gel charging apparatus according tothe present invention is provided with a guide rod 130 a guiding theholder 4 and fixing means 111 fixing the holder 4, and the holder 4 isprovided with a guide hole 30 b through which the guide rod 130 a passesand a fixing part 4 b so inclined that thickness of four comers reducetoward an outward direction to be engaged with the fixing means 111.

[0078] When clamping the holder plate 6 b with respect to the holderplate 6 a with force beyond necessity in the holder 6 on the detectionside, the holder plate 6 b is bent due to the combined thickness of thecapillary columns and the rubber plate. In order to prevent this, it ispreferable to provide a groove in the holder 6 b in fixing parts of theholder plates 6 a and 6 b in response to the thickness of the rubberplate or to insert a spacer between the holder plates 6 a and 6 b. It isalso preferable to prepare the holder plate 6 b while previously warpingthe same opposite to the bent direction. These are effective forpreventing the holder plate 6 b from being bent, and effective formaintaining the force pressing the rubber plate 16 through the clamp 18and maintaining holding force around the center of the rubber plate 16.

[0079] For carriage of the capillary cassette, it is convenient to use acarrier. FIG. 5 shows a capillary cassette comprising a carrier as anexample. (A) is a front elevational view, (B) is a left side elevationalview, and (C) is a top plan view.

[0080] An end of an L-shaped carrier pipe 22 is fixed to a carrierfixture 20 of a holder 4, a holder fixing part 26 is provided on anotherend of the carrier pipe 22, and a holder 6 is fixed to the holder fixingpart 26 by fixing screws 28. The holders 4 and 6 are fixed by thecarrier pipe 22.

[0081] In the embodiment of FIG. 5, the carrier 22 isattachable/detachable, and it is possible to mount the carrier 22 andfix the holders 4 and 6 when carrying the capillary cassette and detachthe same when mounting the capillary cassette to an electrophoreticapparatus. However, a capillary cassette integrated with a carrier maybe prepared to be mounted on an electrophoretic apparatus whilecomprising the carrier.

[0082]FIG. 6 shows a schematic block perspective view of one embodimentof a capillary cassette preparation apparatus, and an example of acapillary cassette preparation method according to the present inventionis described with reference to this.

[0083] The embodiment employs quartz glass capillary columns of 300 μmin outer diameter and 100 μm in inner diameter. As a sample injectionside holder, it employs a sample injection side holder 1 holdingcapillaries in two dimensions of 16 by 24 of 4.5 mm pitches incoincidence with a 384-hole microplate used in sample injection.

[0084] The sample injection side holder 1 is in such a structure that asupporter consisting of a silicone rubber plate is held between twometal plates or resin plates in which holes are provided on positionsmatched with the 384-hole microplate. A needle 3 (e.g., 0.72 mm in outerdiameter and 0.41 mm in inner diameter) linearly moving in a directionperpendicular to the plane of the supporter and passing through thesupporter is provided. The needle 3 has an inner diameter larger thanthe outer diameter of the capillary columns. A chuck 7 serving as afirst capillary column holder chucking prescribed positions from forwardends of the capillary columns and inserting the forward ends into theneedle 3 is provided, and a guide pipe 5 having an inner diameter largerthan the outer diameter of the capillary columns is provided for guidingthe forward ends of the capillary columns chucked by the chuck 7 intothe needle 3. In order to hold the forward end portions of the capillarycolumns made to project from the forward end of the needle 3 in such astate that the needle 3 passes through the supporter, a chuck 11 servingas a second capillary column holder is also provided. Furthermore, anichrome wire coil 13 preparing detection windows on prescribedpositions from the forward ends of the capillary columns is provided.The guide pipe 5, the chuck 7 and the nichrome wire coil 13 move on thesame straight line as the needle 3. Furthermore, a cutter 15 cutting thecapillary columns whose single end sides are fixed to the sampleinjection side holder 1 in a prescribed length and a capillary feed 17drawing out the capillary columns wound on a capillary drum 19 by aconstant length are also provided.

[0085] It is possible to move the needle 3, the guide pipe 5, the chuck7 and the nichrome wire coil 13 reciprocally between a position furtheron the left side of the position where the needle 3 passes through thesupporter of the sample injection side holder 1 and a position of brokenlines.

[0086] The capillary cassette preparation method according to thepresent invention is formed by the following procedure:

[0087] (1) It mounts the sample injection holder 1 on a non-illustratedXY stage holding the sample injection side holder 1 and sliding the sameupward, downward, leftward and rightward in the figure while directingthe plane of the supporter of the sample injection holder 1 in adirection perpendicular to the needle 3, and matches a hole provided onthe sample injection side holder 1 on the moving straight line of theneedle 3. The hole defines a fixing position for the capillary columns.

[0088] (2) In the state where the chuck is on the position of the brokenlines, it extracts a capillary column from the capillary drum 19 usingthe capillary feed 17 by the same length as the capillary columnscomprised in the capillary cassette. The extracted capillary column isstored between the chuck 7 and the capillary feed 17 in a loosenedstate.

[0089] (3) It passes through the supporter in the sample injectionholder 1 by the needle 3.

[0090] (4) It presses the capillary column whose prescribed positionfrom forward end is chucked by the chuck 7 in the left direction, tomake the forward end pass through the supporter through the guide pipe 5and the needle 3. When the needle 3 is clogged with sediment of thesupporter, it pushes out the same with the capillary column. At thistime, the capillary column between the needle 3 and the chuck 7 isprotected by the guide pipe 5, whereby it is possible to prevent thecapillary column from being bent and broken by the force pressing thesediment of the supporter in the needle 3.

[0091] (5) It chucks the forward end of the capillary column passingthrough the needle 3 and projecting on the opposite side of the sampleinjection side holder 1 with the chuck 11.

[0092] (6) It removes the coat of the capillary column on the prescribedposition from the forward end of the capillary column with heat of thenichrome wire coil 13 and prepares a detection window.

[0093] (7) It extracts only the needle 3 from the supporter of thesample injection side holder 1. The capillary column passing through thesupporter is fixed to the sample injection side holder 1 by elasticforce of the supporter acting in a direction closing the hole.

[0094] (8) In a state chucking and holding the forward end of thecapillary column with the chuck 1, it moves the needle 3, the guide pipe5, the chuck 7 and the nichrome wire coil 13 while sliding the same inthe direction of the capillary feed 17 along the capillary column.Thereafter it cuts the capillary column in a prescribed length from theforward end with the cutter 15.

[0095] (9) The cut side (detection side) of the capillary column whoseone end is fixed to the sample injection holder 1 is dropped into twoslit holders 21 a consisting of two planes having parallel clearancesslightly wider than the outer diameter (300 μm) of the capillary column,and aligned in cut order, i.e., the order fixed to the sample injectionside holder 1.

[0096] (10) The XY stage moves so that a next fixing position of thecapillary cassette 1 is located on a moving locus of the needle 3.

[0097] (11) It repeats (2) to (10) 384 times, fixes the capillarycolumns to all fixing positions of the capillary cassette 1, thereafteraligns end surfaces of the aligned capillary columns on the detectionside with the slit holders 21 a, and mounts/fixes the detection sideholder.

[0098] When executing the capillary cassette preparation methodaccording to the present invention, preparation of a capillary cassettecan be simply and quickly performed.

[0099]FIG. 7 and FIG. 8 show block diagrams of one embodiment of acapillary cassette preparation apparatus for more reliably carrying outthe aforementioned capillary cassette preparation method. FIG. 7 is aplan view and FIG. 8 is a front elevational view. The same referencenumerals are assigned to parts playing the same roles as FIG. 6.

[0100] Three struts 25 a, 25 b and 25 c are comprised on a substrate 23in a direction perpendicular to the plane of the substrate 23. A slideplate 27 comprising a rail 27 a sliding a needle unit 3 a, a guide unit5 a, a holder unit 7 a and a detection window preparation unit 13 adescribed later in a direction parallel to the substrate 23 on its sidesurface is fixed over the three struts 25 a, 25 b and 25 c. An XY stageis comprised on the substrate 23 on extension of the longitudinaldirection of the slide plate 27 with its moving direction (XY plane)along a vertical direction. A sample injection side holder 1 having asilicone rubber plate as a supporter is mounted on the XY stage 29 withthe plane of the supporter perpendicular, and it can be moved upward,downward, leftward and rightward. A chuck 11 holding and keeping aforward end of a capillary column is comprised on an opposite side (aside opposite to the slide plate 27) of the XY stage 29 on extension ofthe longitudinal direction of the slide plate 27.

[0101] A needle 3 having an inner diameter larger than the outerdiameter of the capillary column and passing through the silicone rubberplate which is the supporter of the sample injection holder 1 iscomprised in the needle unit 3 a sliding on the rail 27 a comprised onthe slide plate 27. A guide pipe 5 having an inner diameter larger thanthe outer diameter of the capillary column for making the capillarycolumn between the needle unit 3 a and the holder unit 7 a describedlater not bent is comprised in the guide unit 5 a. A chuck 7 chucking aprescribed position from the forward end of the capillary column iscomprised in the holder unit 7 a. A nichrome wire coil 13 burning andremoving a coat of the capillary column is comprised on the detectionwindow preparation unit 13 a, and the capillary column passes throughthe center of the nichrome wire coil 13. A ceramic pipe cutting offtransmission of heat from the nichrome wire coil 13 to the capillarycolumn is provided between the capillary column and the nichrome wirecoil 13 to be movable along the capillary column. Around the strut 25 con the side surface of the slide plate 27, a pipe unit 31 guiding thecapillary column to the holder unit 7 a is fixed so that the capillarycolumn fed from a capillary drum 19 described later becomes linear alongthe slide plate 27.

[0102] The chuck 11, the needle 3, the guide pipe 5, the chuck 7 and thenichrome wire coil 13 are provided on a straight line.

[0103] The capillary drum 19 winding and storing a single long capillarycolumn is comprised on the substrate 23 on the opposite side to the XYstage.

[0104] A cut unit 15 a cutting the capillary column after being held bythe sample injection side holder 1 in a prescribed length by moving acutter upward and downward is comprised on the strut 25 b.

[0105] A detection side cassette preparation unit 21 is provided on thesubstrate 23 between the strut 25 a and the strut 25 b, and two slitholders 21 a consisting of two planes having parallel clearancesslightly wider than the capillary column outer diameter are comprised ona rotary plate 21 b provided in parallel with the substrate 23. Therotary plate 21 b is in contact with the substrate 23 through a fulcrumplate 21 c and a roller 21 d, and the longitudinal direction of therotary plate 21 b is fixed in a direction parallel to the slide plate 27during an operation of fixing the capillary column to the sampleinjection side holder 1, and the clearances of the slit holders 21 a arealso directed to the direction parallel to the slide plate 27, i.e., thedirection of the XY stage 29 at this time. It is in such a structurethat the detection side cassette preparation unit 21 can rotate in theopposite direction to the slide plate 27 by 90° in a direction parallelto the substrate 23 about a non-illustrated rotation axis provided inthe fulcrum plate 21 c in detection side cassette preparation. Byrotating the detection side cassette preparation unit 21 by 90°, thecapillary column enters a state bent by 90° as shown in FIG. 1, and isthereafter fixed to the detection side holder.

[0106]FIG. 9 shows a flowchart of an operation procedure of theaforementioned embodiment, and description of the operation is madeemploying FIG. 7 to FIG. 9.

[0107] It moves the XY stage and matches a position of the sampleinjection side holder 1 for first fixing the capillary column on thestraight line of the chuck 11, the needle 3, the guide pipe 5, the chuck7 and the nichrome wire coil 13. The positions of the needle unit 3 a,the guide pipe 5 a, the holder unit 7 a and the detection windowpreparation unit 13 a shown in FIG. 7 and FIG. 8 are initial states. Inthis state, the forward end of the capillary column wound on thecapillary drum 19 is fed to the pipe unit 31 through a non-illustratedcapillary feed having a motor for motive power. Furthermore, the forwardend is fed into the guide pipe 5 through the nichrome wire coil 13 andthe chuck 7, and the capillary column is held and kept by the chuck 7when the forward end goes out from the guide pipe 5. While the nichromewire coil 13 is fed with a current and heated, the ceramic pipeintervenes between the nichrome wire coil 13 and the capillary column sothat the heat of the nichrome wire coil 13 is not transmitted to thecapillary column.

[0108] (STEP 1) It moves the needle unit 3 a and the guide unit 5 a inthe left direction with a knob comprised on the guide unit 5 a. It movesthe needle unit 3 a along the rail 27 a leftward until the needle 3sticks into and passes through silicon rubber which is the supporter ofthe sample injection side holder 1 set on the XY stage 29. At thisstage, the holder unit 7 a and the detection window preparation unit 13a follow the guide unit 5 a while keeping a prescribed space and movealong the rail 27 a. The capillary column is held by the chuck 7 of theholder unit 7 a, to be pulled in the left direction at the same time.

[0109] (STEP 2) It moves the guide unit 5 a further leftward to bringthe same into contact with the needle unit 3 a, guides the forward endof the capillary column going out from the guide pipe 5 into the needle3, makes the forward end of the capillary column project from theforward end of the needle 3, and pushes out sediment of the siliconerubber in the needle 3.

[0110] (STEP 3) It moves the holder unit 7 a further leftward and makesthe forward end of the capillary column further project from the forwardend of the needle 3. At this stage, the ceramic pipe between thenichrome wire coil 13 of the detection window preparation unit 13 a andthe capillary column is moved from the position of the nichrome wirecoil 13 on a set time, so that the nichrome wire coil 13 heats a coat ona prescribed position from the forward end of the capillary column andstarts preparation of a detection window. Since it requires time forpreparation of the detection window, it performs operations of STEP 4and STEP 5 in parallel.

[0111] (STEP 4) It holds the forward end of the capillary columnprojecting from the forward end of the needle 3 with the chuck 11.

[0112] (STEP 5) It moves the needle unit 3 a and the guide unit 5 a inthe right direction until the needle 3 comes out of the silicone rubber,and reduces the space between the same and the holder unit 7 a. At thisstage, the capillary column is held by the chucks 7 and 11, remains inthe state passing through the silicone rubber, and is fixed to thesample injection side holder 1 by elastic force of the silicone rubber.

[0113] (STEP 6) It opens the chuck 7, moves the needle unit 3 a, theguide unit 5 a, the holder unit 7 a and the detection window preparationunit 13 a rightward, and returns the holder unit 7 a and the detectionwindow preparation unit 13 a to the positions of the initial states. Atthis stage, the space between the guide unit 5 a and the holder unit 7 ais shorter than the initial state.

[0114] (STEP 7) The holder unit 7 a closes the chuck 7 on the positionin the initial state, and holds the capillary column.

[0115] (STEP 8) It moves the cut unit 15 upward, and cuts the capillarycolumn with the comprised cutter. The cut portion defines the forwardend of a next capillary column. An end of the capillary column cut inthe prescribed length is fixed to the sample injection side holder 1,and another end is dropped in the clearance of the slit holders 21 a.The capillary column fixed to the sample injection side holder 1 issuccessively dropped in the clearance of the slit holders 21 a, andplanarly arranged in a line.

[0116] (STEP 9) It moves the needle unit 3 a and the guide unit 5 aleftward and returns the space between the guide unit 5 a and the holderunit 7 a to the position in the initial state. Thus, the needle unit 3a, the guide unit 5 a, the holder unit 7 a and the detection windowpreparation unit 13 a return to the initial positions.

[0117] (STEP 10) It opens the chuck 11 and releases the chucked forwardend of the capillary column. Thereafter it moves the XY stage and thenmatches the position of the sample injection side holder 1 for fixingthe capillary column on the straight line of the chuck 11, the needle 3,the guide pipe 5, the chuck 7 and the nichrome wire coil 13. Whenfixation of all capillary columns ends, it advances to STEP 11,otherwise returns to STEP 1.

[0118] (STEP 11) It moves the XY stage 29 to the lowermost position forpreparing a capillary cassette, and moves the sample injection sideholder 1 to a prescribed position.

[0119] (STEP 12) In such a state that 384 capillary columns are arrangedin a line on a plane in the two slit holders 21 a, it rotates the rotaryplate 21 b of the detection side cassette preparation unit 21 in theopposite direction to the slide plate 27 with a roller 4 d by 90° abouta non-illustrated rotation axis provided in the fulcrum plate 21 c. Atthis time, each capillary column is bent by 90°.

[0120] (STEP 13) It rectilinearly aligns end surfaces of the 384capillary columns going out from the slit holders 21 a with a prescribedjig (angle). At this time, a detection window provided in each capillarycolumn is positioned between the two slit holders 21 a.

[0121] (STEP 14) It holds and fixes the 384 capillary columns withclamps comprised in the slit holders 21 a holding the capillary columnspositioned in the clearances of the slit holders 21 a.

[0122] (STEP 15) It matches the detection side cassette along betweenthe two slit holders 21 a and screws the same.

[0123] By operating a lever in accordance with the aforementionedprocedure, capillary columns of a predetermined length are aligned inset order, so that a capillary cassette can be prepared.

[0124] While a manual example has been shown as to the operation in theembodiment, it is preferable to make automatization with a drivingsource such as a motor or an air cylinder. Thus, preparation of thecapillary cassette can be further simply performed without failure.

[0125] While the capillary columns have been charged one by one in theembodiment, it is preferable to arrange a plurality of, for example, 16needles, and capillary columns and simultaneously move the same. Thus,preparation of the capillary cassette in a shorter time can beperformed.

[0126] Another embodiment forming a detection window on the position ofa detected portion 2 c in a capillary cassette 2 is described withreference to FIG. 10.

[0127] (A) After fixing sample injection sides of 384 capillary columns102 not subjected to coat removal with a holder, it aligns other endsides, i.e., detection sides in a line on the planar of a flat plate 204in a close contact manner. A slit longer than the arrangement width ofthe capillary columns 102 is opened in the flat plate 204 in order toprevent the capillary columns 102 from adhering to the flat plate 204 bymelting of coats through a coat removal operation, and it performspositioning so that a detection window formation planned region 102 acomes onto the slit

[0128] (B) It further overlaps another flat plate 206 on the capillarycolumns 102. A slot longer than the arrangement width of the capillarycolumns 102 is opened also in the flat plate 206, and it performspositioning so that the slot comes to the detection window formationplanned region 102. Thus, it holds and fixes the capillary columns 102between the two flat plates 204 and 206.

[0129] (C) It feeds a current to a nichrome wire coil 208 to bum thecoats and makes it heat red. The nichrome wire coil 208 is that windinga 40 cm long of a nichrome wire of 0.5 mm in diameter, wound into theform of a coil of about 3 mm in diameter and 2 cm in length. Theenergization quantity is, for example, 5A.

[0130] It brings the red-heating nichrome wire coil 208 into contactwith the capillary columns 102 while directing the same to an endportion of the detection window formation planned region 102 a throughthe slit of the flat plate 206 so that the longitudinal direction of thenichrome wire coil 108 is along the arrangement direction (the directionperpendicular to the longitudinal direction of the capillary columns102) of the capillary columns 102.

[0131] (D) It brings the nichrome wire coil 208 into contact with thecapillary columns 102 for about 10 seconds, bums and removes the coatson the positions, and thereafter moves the nichrome wire coil 208 alongthe slot of the flat plate 208 toward the other end portion direction ofthe detection window formation planned region 102 a while partiallyoverlapping portions for bringing the nichrome wire coil 208 intocontact so that the coats do not remain on the detection windowformation planned region 102 a. At this time, coats on surfaces oppositeto the surfaces with which the nichrome wire coil 208 comes into contactare also burned and removed.

[0132] When the coats of the respective capillary columns 102 melted inpositions separate from the nichrome wire coil 208 in burning are cooledand solidified, the adjacent capillary columns 102 are fused to eachother by the coats once melted. Thus, the overall plurality capillarycolumns 102 arranged in a line form a flat cable.

[0133] While the embodiment has used the coiled nichrome wire as a meansof heating, a long and narrow ceramic heater or the like can also beused as the heating means. When the number of the capillary columnsfixed to the holder is large, the width when planarly aligning thecapillary columns in a line becomes large. When lengthening the nichromewire coil for reducing the time required for removal of the coats, itsstrength weakens in such a manner that the nichrome wire coil is bentand deformed while preparing a detection window by pressing the sameagainst the capillary columns and cannot indeed come into contact withthe planarly aligned capillary columns, and it may become necessary torepeatedly perform the burning operation over and over. When using aceramic heater longer than the width when planarly aligning thecapillary columns in a line in such a case, it is possible to form adetection window at once and hence the detection window forming time canbe reduced.

[0134]FIG. 11, FIG. 12 and FIG. 13 show schematic block diagrams of oneembodiment of a capillary gel charging apparatus according to thepresent invention. FIG. 11 is a front elevational view showing a closingmeans part and a fixing means part in a partially fragmented manner,FIG. 12 is a plan view showing an upper portion of a pump means storagebox and a timer unit set therein in a partially fragmented manner, andFIG. 13 is a sectional view showing the pump means storage box in apartially fragmented manner, showing the same while cutting the samealong an A-A line position of FIG. 12.

[0135] A chamber 103 in which the sample injection side holder 4 shownin FIG. 2, FIG. 3 and FIG. 4 is fixed to form a closed space, a pumpmeans storage box 105 comprising pump means pressurizing ordecompressing the chamber 103 therein, detection side holder fixingmeans 107 and detection side gel vessel fixing means 129 are comprisedon a substrate 101.

[0136] The chamber 103 is formed by 170 mm×170 mm square acrylic plates103 a and 103 c of 10 mm in thickness for closing an upper portion and abottom portion, and an acrylic pipe 103 b of 165 mm in outer diameterand 145 mm in inner diameter for closing side portions. Clearancesbetween both ends of the acrylic pipe 103 b and the acrylic plates 103 aand 103 c are sealed through a silicone rubber packing. An opening 113of a rectangular hole of 80 mm by 115 mm is provided on the acrylicplate 103, and the sample injection side holder 4 is mounted thereon. Agroove is dug around the rectangular hole opening 113, and an annularsilicone sponge 113 a having a circular section is fit into the groove.About half the silicone sponge 113 a goes out from the surface of theacrylic plate 103 a, to seal a clearance between the acrylic plate 103 aand the sample injection side holder 4. Four clamps 111 for fixing thesample injection side holder 4 to the acrylic plate 103 a are comprisedon the acrylic plate 103 a and engage with fixing parts 4 d on fourcorners of the sample injection holder 4, so that the holder 4 ispressed against the sponge 113 a and closes the space of the chamber 103by tightening the clamps 111.

[0137] Two joints are provided on a side surface (cylindrical surface)of the acrylic pipe 103, and a pump 119 pressurizing or decompressingthe chamber 103 is connected to one joint 115. An electromagnetic valve121 releasing the pressure in the pressurized or decompressed chamber103 to the atmospheric pressure through a needle valve 117 is connectedto the other joint, and its return pressure speed is adjusted by theneedle valve 117.

[0138] In the pump means storage box 105, a timer 123 a controlling aworking time of the pump 119 and a timer 123 b controlling a workingtime of the electromagnetic valve 121 are stored in addition to the pump119 and the electromagnetic valve 121.

[0139] A detection side gel vessel 129 storing a gel solution wheninhaling the gel solution from the detection side is comprised in thedetection side gel vessel fixing means 129. It uses a polyacrylamidesolution of 5% T and 5% C, for example, for the gel solution.

[0140] The operation shall now be described. The embodiment has employedthe capillary cassette comprising the 384 capillary columns 102 shown inFIG. 2, FIG. 3 and FIG. 4.

[0141] Sample injection sides of the 384 capillary columns 102 arearranged in two dimensions of 16 by 24 of 4.5 mm pitches in coincidencewith a commercially available microplate by the sample injection holder4 and fixed with excellent airtightness, and detection sides thereof areplanarly closely fixed in a line by the detection side holder 6.

[0142] When performing gel charging by decompressing the chamber 3, itstores the gel solution in a detection side gel vessel 129 a. It fixesthe fixing parts 4 d on the four corners of the detection side holder 4by the four clamps 111 provided on the acrylic plate 103 a so that theforward ends on the detection sides of the capillary columns 102 aredipped into the gel solution stored in the detection side gel vessel 129a, presses the sample injection side holder 4 against the siliconesponge 113 a by tightening the clamps 111, and closely fixes the same tothe acrylic plate 103 a. It decompresses the chamber 103 with the pump119 for decompressing the inner parts of the capillary columns 102, andinhales the gel solution from the forward ends on the detection sides.The gel solution is charged into the capillary columns 102, and it ispreferable to comprise a non-illustrated vessel in the chamber 103 toreceive an excess amount of gel solution overflowing from the sampleinjection side.

[0143] When performing gel charging by pressurizing the inner part ofthe chamber 103, it comprises a non-illustrated vessel storing the gelsolution and vacates the vessel 129 a. It adjusts the length of thecapillary columns projecting from the holder 4 and the height of thevessel storing the gel solution so that the forward ends of allcapillary columns on the sample injection side are dipped into the gelsolution when fixing the fixing parts 4 d on the four corners of thesample injection side holder 4 with the four clamps 111 provided on theacrylic plate 103 a. It fixes the detection side holder 6 to thedetection side holder fixing means 107. In this case, the sampleinjection side holder 4 is pressed against the silicone sponge 113 bytightening the clamps 111 and closely fixed to the acrylic plate 103 a,so that the space inside the chamber 103 enclosed with the sampleinjection side holder 4, the acrylic plates 103 a and 103 b and theacrylic pipe 103 is closed. It pressurizes the chamber 103 with the pump119 and pushes the gel solution into the capillary columns by thepressure. The gel solution is charged into the capillary columns 102,and it receives an excess amount of gel solution overflowing from thedetection side by the detection side gel vessel 129 a.

[0144] Both in the case of performing decompression and, in the case ofperforming pressurization, it times the time when the gel solution ischarged into all capillary columns 102 and previously sets the workingtimes of the pump 119 and the electromagnetic valve 121 with the timers123 a and 123 b. When charging of the gel solution is started by a RUNbutton, the pump 119 starts working and the chamber 103 is decompressedor pressurized. Continuing the decompression or the pressurization bythe time set at the timer 123 a, the pump 119 stops at a prescribedtime. The inner part of the chamber 103 is still decompressed orpressurized even if the pump 119 stops, whereby the electromagneticvalve 121 opens when the time set at the timer 123 b elapses so that thepressure in the chamber 103 is released to the atmosphere through theneedle valve 117. At this time, the speed at which the internal pressureof the chamber 103 returns is adjusted by the needle valve 117.

[0145] Decompression-pressurization in the chamber 103 can be selecteddepending on to which one of an exhaust port and a suction port of thepump 119 a pipe connected to the joint 115 is connected.

[0146] In order to prevent air from being mixed into the gel chargedinto the capillary columns since the degree of decompression orpressurization of the chamber 103 is large, it is preferable to controlthe degree of decompression or pressurization. This control can beperformed by a throttle comprised in the needle valve 117 while openingthe electromagnetic valve 121 also when the pump 119 operates by thetimer 123 a. In this case, the electromagnetic valve 121 temporarilycloses when the set time of the timer 123 a elapses, and opens againafter a lapse of the time set in the timer 123 b to release the chamber103 to the atmosphere.

[0147] While it is of a pump storing type in the embodiment, it ispreferable to render a pressure generation source outside and connectthe same to, for example, a high pressure cylinder or a vacuum pump,when the charged gel solution is that having high viscosity such as aflow gel, and it is necessary to generate a high pressure. Whenemploying a high pressure cylinder, it is preferable to provide a newelectromagnetic valve, control the same with a timer, and introducecompressed gas in the cylinder into the chamber.

[0148] While the embodiment has used a diaphragm pump for enablingswitching of decompression-exhaust, a pump of only suction or onlyexhaust may be employed.

INDUSTRIAL AVAILABILITY

[0149] As described above, the capillary cassette according to thepresent invention is suitable for use as a migration part of amulti-capillary electrophoretic apparatus used for separating/analyzingnucleic acid, protein, peptide, sugar or the like, and particularlysuitable for employment for analysis of the base sequence of DNA.

1. In a capillary cassette two-dimensionally arranging and fixing aplurality of capillary columns used in a multi-capillary electrophoreticapparatus on a sample injection side with a holder while aligning thesame in a line on a plane on a detection side, a capillary cassettecharacterized in that said holder comprises a rubber plate, and endportions of said plurality of columns pass through the rubber plate oneby one and are fixed by elastic force of the rubber plate.
 2. Thecapillary cassette in accordance with claim 1, comprising another holderpressing and fixing said plurality of capillary columns arranged in aline on the plane from at least single surface sides on detection sidesof said plurality of capillary columns.
 3. The capillary cassette inaccordance with claim 2, wherein said holder on detection sidescomprises, as a means of fixing the plurality of capillary columnsarranged in a line on the plane through said rubber plate, clampingmeans fixing the same on both side portions of a capillary columnarrangement while pressing and fixing the same also on at least twoportions of an intermediate part.
 4. A capillary cassette preparationmethod characterized in passing a supporter of a prescribed fixingposition of capillary column fixing means having said supporterconsisting of a rubber plate with a needle having an inner diameterlarger than a capillary column from its vertical direction, guiding thecapillary column into said needle and making the same pass through saidsupporter, thereafter extracting only said needle thereby holding andfixing the capillary column by said supporter and repeating thisoperation while changing the fixing position on the supporter therebysuccessively two-dimensionally arranging and fixing a plurality ofcapillary columns.
 5. A capillary cassette characterized in that aplurality of capillary columns used in a multi-capillary electrophoreticapparatus are fixed by a holder on a sample injection side and planarlyarranged in a line on a detection side, coats of the capillary columnsare removed and a strip-shaped detection window extending in a capillarycolumn arrangement direction is formed on a capillary column arrangementon the detection side, while the capillary columns adhere to each other,coats fuse to each other and the capillary column arrangement isintegrated around the detection window.
 6. In a method of manufacturingsuch a capillary cassette that a plurality of capillary columns used ina multi-capillary electrophoretic apparatus are fixed by a holder on asample injection side and planarly arranged in a line on a detectionside and a detection window in which coats of the capillary columns areremoved is formed on the detection side, a method of manufacturing acapillary cassette characterized in forming the detection window whileincluding the following steps (A) and (B): (A) a step of fixing sampleinjection sides of the capillary columns with the holder and thereafterplanarly arranging the capillary columns in a line in close contact witheach other; and (B) a step of bringing heating means having a length fora plurality of capillary column outer diameters into contact with orapproximating the same to a detection window formation planned region ofa held capillary column arrangement for removing coats of the pluralityof capillary columns while melting coats around regions where the coatsare removed and thereafter solidifying the same for fusing coats ofadjacent capillary columns to each other.
 7. The method of manufacturinga capillary cassette in accordance with claim 5, wherein the length ofsaid heating means is shorter than the width of the capillary columnarrangement on the detection side, and it repeats said step (B) aplurality of times in the width direction of said capillary columnarrangement.
 8. A gel charging apparatus for charging a gel sewing as amedium in which a sample is electrophoresed into a plurality ofcapillary columns mounted on a multi-capillary electrophoreticapparatus, wherein end portions on sample injection sides of thesecapillary columns pass through a holding member of a holder and aretwo-dimensionally arranged and fixed while keeping airtightness betweenthe same and the holding member to form a capillary cassette, and thisgel charging apparatus comprises a chamber provided with an opening onits upper surface so that closure means inserting end portions on thesample, injection sides of said capillary columns inside for closing theopening with said holder is provided in the opening, and is furtherprovided with an exhaust port and a release port to the atmosphere; agel vessel storing a gel solution so that end portions of said capillarycolumns on opposite sides to the sample injection sides are dippedtherein; exhaust means provided on the exhaust port of said chamber; anda switching valve provided on said release port.
 9. A gel chargingapparatus for charging a gel becoming a medium in which a sample iselectrophoresed in a plurality of capillary columns mounted on amulti-capillary electrophoretic apparatus, wherein end portions onsample injection sides of these capillary columns pass through a holdingmember of a holder and are two-dimensionally arranged and fixed whilekeeping airtightness between the same and the holding member to form acapillary cassette, and this gel charging apparatus is characterized incomprising a chamber provided with an opening on its upper surface sothat closure means inserting end portions on the sample injection sidesof said capillary columns in its inner side for closing the opening withsaid holder is provided in the opening, stores therein a gel solution onpositions where the end portions on the sample injection sides of saidcapillary columns are dipped in the state where said opening is closedwith said holder, and is further provided with a pressurization port anda release port to the atmosphere; pressurization means provided on thepressurization port of said chamber; and a switching valve provided onsaid release port.
 10. The gel charging apparatus in accordance withclaim 8, wherein a flow control valve controlling the flow rate isprovided between said release port and said switching valve.
 11. The gelcharging apparatus in accordance with claim 9, wherein a flow controlvalve controlling the flow rate is provided between said release portand said switching valve.